OBJECTIVE To find out the situation of the use of preventive application of antimicrobial in orthopedic perioperative patients and to evaluate the rationality of use. METHODS To make a survey of the use of antimicrobials in orthopedic perioperative patients during Apr-Jun 2004 and to evaluate the kinds,frequency,combined administration,course,effectiveness,etc. RESULTS 99.2% patients had received antimicrobials involving 14 kinds.62.8% from them used one.34.5% used two and 2.7% used three. CONCLUSIONS It is necessary to strengthen the management of preventive antimicrobial in orthopedic patients during perioperative period.

Objective To select the optimum conditions of Echinococcosis ELISA kit and kit preparation by using orthogonal experiments,then to test their detection performance.Methods Taking the absorbance of the samples as the indicator,orthogonal experiments of three factors four levels were taken to optimize the conditions of hydatid disease detection ELISA kit.Through L16 (43)orthogonal experiments,the concentration of the antigen(A),the dilution multiple of the sample (B) and the dilution multiple of the enzyme labeled antigen (C) were studied.The specimen were provided by The First Affiliated Hospital of Xinjiang Medical University specimen library collected between March 2011 and June 2013,including 36 sera confirmed alveolar Echinococcus by the gold standard,56 sera confirmed Echinococcus granulosus disease by the gold standard and 72 sera as the control group(including healthy people,cirrhosis,hepatitis,etc).Detecting sensitivity and specificity were compared using F test for statistical analysis.Results Orthogonal design showed the size proportion of three factors of echinococcosis ELISA kit:antibody dilution ＞ sample dilution ＞ antigen coating concentration.The optimal preparation conditions were A1 B2 C4,that was,the concentration of antigen was 1 mg/L,sample dilution of 1∶ 100,dilution of secondary antibody of 1∶160 000.Through the 3-factor and 4-level orthogonal design using F-test analysis,the fact of the concentration of the kit antigen(A),F =1.181,P =0.393; the fact of the dilution multiple of the sample (B),F =2.544,P =0.152,the fact of enzyme labeled antigen dilution (C) F =2.544,P =0.039.The sensitivity of the kit to Echinococcus granulosus disease and alveolar Echinococcus were 83％ (30/36),and 91％ (51/56),respectively; the specificity was 91％ (29/32) and 83％ (33/40); the misdiagnosis rate of 17％ (6/36) and 9％ (5/56) ; the misdiagnosis rate 9％ (3/32) and 18％ (7/40) ; positive likelihood ratio 8.86％ (83/9) and 5.20％ (91/18) ; negative likelihood ratio of 0.18％ (17/91) and 0.11％ (9/83).Conclusion Orthogonal design is a good method that can find out optimal conditions of preparation for Echinococcosis ELISA kit and improve the test performance.

Objective To clarify the actions of tranilast,an anti allergic drug,on chronic constriction mononeuropathy through an experiment using an animal model according to the method of Bennett et al. Methods 36 rats were divided into 2 groups: one was treated with tranilast(200mg/kg?d-1,p.o every experimental day),the other was controlled with solvent only. Chronological changes of heat evoked withdrawal latencies on a hot plate,nerve coducting velocitics, and histopathological changes were compared between the two groups from one to four weeks after loose ligation in the sciatic nerve. Results The changes of the withdrawal latency and nerve conducting velocities in the tranilast treatment group were significantly longer than those in the control. The rats without tranilast treatment showed inflammation in and around the constricted nerve bundles,axonal degeneration,phagocytes invasion and interstitial edematous changes at 7 days,numerous axonal sprouts and remyelination at 14 days,and regeneration in the nerve bundles at 28 days. In contrast,the rats treated with tranilast showed less inflammation or nerve fiber degeneration,and better regeneration than those of the control. Conclusion The actions of tranilast appear to have beneficial effects on prophylaxis of a chronic constriction mononeuropathy and on preservation of the nerve functions in rats.

HCV NS5B, acting as a RNA dependent replication enzyme,has emerged as an attractive protein used as a target for screenig of drugs against HCV NS5B, and plays an important role in HCV replication. In the report the gene expression of NS5B in E.coli was investigated. PCR was performed to gain the gene of HCV NS5B from plasmid pBRTM/HCV 1 which contains whole sequence of HCV, and the truncated NS5B gene containing no hydrophobic domain was cloned into pGEM Teasy vector. The gene of the truncated NS5B was cut from pGEM Teasy vector and cloned into E.coli expression plasmid pET 21b, then pET 21b NS5B was transfected into E.coli cells. The protein E.coli lysates were purified and analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS PAGE) and Western blotting assay. The RdRp activity of NS5B was examined by scintillation proximity assay (SPA). The truncated NS5B gene was successfully cloned into pET 21b. The results of SDS PAGE and Western blotting assay showed: ①the molecular weight of the expressed product was about 68 000 D, ②The truncated NS5B protein was existed in media of E.coli cells, ③The activity of NS5BDCT21 His from HCV 1b amounted to 6 900 cpm (total incorporation of approximately). These findings suggest that soluble NS5B can be successfully expressed in E.coli and could secret into media.

Objective:To study the optimal extraction process of Yinxiejing. Methods: An orthogonal experimental method was used with reflux time, reflux times, ethanol concentration and volume as the influencing factors, and emodin and osthole content as the indices. Results:The optimal extraction conditions for Yinxiejing were as follows:refluxing twice with 2 h every time using 8-fold 80%ethanol. Conclusion:The optimum process is stable and feasible.

Objective To investigate the effect of mouse double minute 2 (MDM2) mRNA ASON and mismatched oligonucleotide (ASONM) radiolabeled with 99Tcm on target gene expression in LNCaP cells.Methods The ASON and ASONM targeted to MDM2 mRNA were synthesized and radiolabeled by 99Tcm with the bifunctional chelator of HYNIC.The labeling efficiency,radiochemical purity,stability and molecular hybridization activity were investigated.The different concentrations of 99Tcm-HYNIC-ASON (0,100,500 nmol/L) and 99Tcm-HYNIC-ASONM (500 nmol/L) coated with lipofectamin 2000 were incubated with prostate cancer cells for 24 h,then RT-PCR and Western blot were carried out to assay the MDM2,p53 mRNA and the corresponding protein level.The variables of RT-PCR and Western blot were analyzed using one-way analysis of variance and q test.Results The labeling efficiency of ASON and ASONM were (65.15± 2.05)％ (n=5) and (64.93±2.18)％ (n=5),respectively.The radiochemical purity were both more than 90％.99Tcm-HYNIC-ASON had a good stability and could hybridize to the sense oligonucleotide (SON).The contents of MDM2 mRNA in 0,100,500 nmol/L 99Tcm-HYNIC-ASON and 500 nmol/L 99Tcm-HYNIC-ASONM groups were 0.458±0.035,0.250±0.026,0.174±0.032,0.463±0.033,respectively,and there were significant differences between each 2 groups except between 0 nmol/L 99Tcm-HYNIC-ASON and 500 nmol/L 99Tcm-HYNIC-ASONM groups (F=33.69,q =24.32-91.45,all P＜0.01).The average density of MDM2 protein in the 4 groups were 90.712±3.042,71.218±2.915,32.775±3.062,88.121±2.710,respectively (F=235.93,q=6.43-19.14,all P＜0.01; except 0 nmol/L99Tcm-HYNIC-ASON and 500 nmol/L 99Tcm-HYNIC-ASONM).The contents of p53 mRNA in the 4 groups were 0.185±0.046,0.203±0.040,0.213±0.027,0.163±0.049,respectively(F =2.18,P＞ 0.05).The average density of p53 protein was 33.865 ± 2.213,70.445±2.180,99.025±3.012,38.351±3.271,respectively (F=53.98,q =3.32-6.74,all P＜0.01 ; except 0 nmol/L 99Tcm-HYNIC-ASON and 500 nmol/L 99Tcm-HYNIC-ASONM).Conclusions The MDM2 antisense probe can accumulate in the prostate cancer cells,and specially hybridize to the MDM2 mRNA and inhibit target gene expression.This novel molecular probe has a promising potential for the diagnosis of prostate cancer at gene level.

Objective:To study the effects of rat interleukin-10 (rIL-10) gene treatment on the expression of collagen , matrix metalloproteinase 13(MMP13) and their specific inhibitors the tissue inhibitor of metalloproteinase 1(TIMP1) in porcine serum in-duced liver fibrosis rats then to explore the anti-fibrotic effect of rL-10.Methods:Thirty SD rats were divided into normal control and fibrosis model group.Normal control group (group C) was intraperitoneally injected with 0.5 ml normal sodium twice a week for 8 week, while the fibrosis model group was injected with equal volume of pig serum for 8 week.At the beginning of the 5th week, fibrosis model group was further randomly divided into a fibrosis model subgroup ( group M ) , rIL-10 gene treatment subgroup ( group I ) and empty vector control subgroup(group P).Rats in group C and M were injected with Ringer’s solution as a reagent control via the tail vein weekly, rats in group I were injected with the rIL-10 plasmid pcDNA3-rIL-10, and rats in group P were injected with empty vector pcDNA3.All rats were sacrificed at the end of 8th week, and the liver tissue samples were collected to observe deposition of collegan in liver tissue by sirius red staining and detected the expression of MMP 13 and TIMP1 in the liver tissue by SP immunohistochemistry .Re-sults:Sirius red staining showed that the area of the collegan deposition was dramatically increased in fibrosis model subgroup and emp -ty vector control subgroup compared with the normal control group , and the area of the collagen deposition was dramatically decreased in rIL-10 gene treatment subgroup compared with the fibrosis model and empty vector control subgroup .Immunohistochemistry analysis showed that the expression of MMP 13 and TIMP1 in fibrosis model subgroup and empty vector control subgroup was significantly higher than the normal control group , but compared with normal control group , expression of MMP13 was significantly increased and expres-sion of TIMP1 was significantly decreased in rIL-10 gene treatment subgroup .Compared with fibrosis model subgroup and empty vector control subgroup, the expression of MMP13 and TIMP1 was dramatically decreased in rIL-10 gene treatment subgroup.Conclusion:rIL-10 gene treatment attenuates the area of collagen deposition in liver fibrosis rats associated with downregulation of TIMP 1.

Objective To investigate the clinical effects of preoperative eye position training on femtosecond laser-assisted laser in situ keratomileusis(FL-LASIK)? Methods One hundred and sixty-eight myopia patients(328 eyes)scheduled for selective FL-LASIK were randomly divided into the experiment group of 85 cases(166 eyes)and the control group of 83 cases(162 eyes)?The patients in the control group were given the routine preoperative education and the patients in the experiment group received the preoperative intervention of eye position besides routine preoperative education? The two groups were compared in terms of intraoperative changes of eye position,corneal topography and the postoperative visual acuity? Results The frequency of eye position adjustment and machine stopping in the experiment group was significantly smaller than that of the control group(P < 0?001)? But no significant differences were found between the two groups in corneal topography and the postoperative visual acuity(P > 0?05)? Conclusion Preoperative training of eye position may maintain ideal eye position and reduce the frequency of downtime due to eye position adjustment during FL-LASIK,which may ensure the successful completion of FL-LASIK?

Objective To study the prevention and treatment of severe complications of percutaneous radiofrequency ablation(PRFA)for hepatic malignancy. Methods A series of 939 patients with primary hepatic carcinoma or hepatic metastasis confirmed by pathological examinations or clinical manifestations underwent 1098 treatments of PRFA between January 2006 and December 2009. All the patients were followed up to study the short-or long-term complications related to PRFA. Results Complications developed in 9 patients: bile duct injury (4 patients), hemothorax (2 patients), and intra-abdominal hemorrhage (3 patients). The incidence of complication was 0.82% (9/1098) and the complication-related mortality was 11.1% (1/9). Conclusions Although PRFA which is minimally invasive, is a safe and effective treatment, there were still risks for this procedure, especially when the tumor is located at the portahepatic region or the patient has coagulopathy. Some serious complications can be prevented. It is important to observe the strict indications for RFA and to carry out the procedure carefully. Early detection of complication is important.

Objective To study the effect of the serum testosterone/estradiol ratio (T/E2) alteration on sperm defect and fertility. Methods The testosterone, estradiol, FSH, LH, PRL,sperm parameters and sperm morphology of 90 men were analyzed and the T/E2 and multiple anomalies index (MAI) were calculated. The patients were divided into 3 groups; T/E2≤10 (Ⅰ), T/E2＞10 (Ⅱ), and T/E2＞20 (Ⅲ). Results The sperm concentration and motility among the 3 groups were not significantly different (P＞0. 05). The percentages of the sperm whose head, neck and tail were abnormal declined gradually with the increase of the T/E2 in serum. The percentage of sperm head defeet of group Ⅰ was significantly higher than that of group Ⅲ (t=2. 482, P=0. 016) and that of sperm neck defect of groups Ⅰ and Ⅱ were significantly higher than that of group Ⅲ (t=4.113, 2. 050, P=0.000, 0. 046, respectively). The percentage of sperm tail defect among 3 groups was significantly different (t=2. 722, 3. 996, 3. 110, P=0. 008, 0. 000, 0. 003, respectively). The SDI of group Ⅱ was significantly higher than that of group Ⅰ (t= -2. 293, P= 0. 025). But the TZI increased gradually with the increase of the serum T/E2 and the TZI of groups Ⅱ and Ⅲ were significantly higher than that of group Ⅰ (t=2. 285, 2. 727, P=0. 025, 0. 009, respectively). The percentage of the men in group Ⅰ whose partners became pregnant was 29. 5% and those of groups Ⅱ and Ⅲ were 50% and 42.9%, respectively. Although the percentage among three groups was not different statistically (x2 = 3. 285, 0. 854, 0. 199, P= 0. 070, 0. 355, 0. 655, respectively), the relative risks of groups Ⅱ and Ⅲ were 2.4 and 1.8 times of that of group Ⅰ. There were 25, 15, 7 cases of idiopathic infertility among the 3 groups, respectively. The relative risk of I group was 1.5 and 1.3 times of that of Ⅱ and Ⅲ groups. The correlation analysis showed that the T/E2 in serum had significantly negative correlation with the percent of the sperm head or neck or tail defects (r= -0. 209, -0. 316 and -0. 335,respectively and P= 0. 048, 0. 002 and 0. 001, respectively). Conclusions The decrease of T/E2 in serum was correlative with the decrease of fertility probability, but it did not alter the sperm density and the sperm motility. It showed that the level of the T/E2 in serum was important for spermatogenesis and sperm fertilizing capability.